quant it picogreen assay kit  (Thermo Fisher)


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    Name:
    Quant-iT PicoGreen dsDNA Assay Kit
    Description:
    Using the PicoGreen dsDNA quantitation assay, you can selectively detect as little as 25 pg/ml of dsDNA in the presence of ssDNA, RNA, and free nucleotides. The assay is linear over three orders of magnitude and has little sequence dependence, allowing you to accurately measure DNA from many sources, including genomic DNA, viral DNA, miniprep DNA, or PCR amplification products.PicoGreen dsDNA Quantitation Reagents are:• Orders of magnitude more sensitive than UV absorbance readings, saving on precious sample (Figure 1)• Specific for dsDNA in the presence of equimolar amounts of RNA (Figure 2)• Easy to use-just add the dye to sample, wait 5 minutes, and read• Suitable for use in 96- and 384-well plate formats• Compatible with most fluorescence-based microplate readers and fluorometers and FluorescenceApplications: The PicoGreen dsDNA Quantitation Reagent and Kits are ideal for PCR-based assays, microarray samples, DNA damage assays, enzyme activity assays, genomic DNA quantitation, measuring dsDNA in complex mixtures, and viral DNA quantitation.
    Catalog Number:
    P11496
    Price:
    None
    Applications:
    DNA & RNA Purification & Analysis|DNA Quantitation|Nucleic Acid Quantitation
    Size:
    10 x 100 µL kit
    Category:
    Kits and Assays, DNA⁄RNA Quantitation Assay Kits
    Score:
    85
    Buy from Supplier


    Structured Review

    Thermo Fisher quant it picogreen assay kit
    Cellular proliferation and viability on the discs. ( A ) <t>Picogreen</t> assay. Proliferation of MC3T3-E1 cells on titanium (Ti) or zirconia (Zr) discs on days 1, 4, and 7. ( B ) CCK-8 assay. Cell viability was tested on days 1, 4, and 7 under the same conditions
    Using the PicoGreen dsDNA quantitation assay, you can selectively detect as little as 25 pg/ml of dsDNA in the presence of ssDNA, RNA, and free nucleotides. The assay is linear over three orders of magnitude and has little sequence dependence, allowing you to accurately measure DNA from many sources, including genomic DNA, viral DNA, miniprep DNA, or PCR amplification products.PicoGreen dsDNA Quantitation Reagents are:• Orders of magnitude more sensitive than UV absorbance readings, saving on precious sample (Figure 1)• Specific for dsDNA in the presence of equimolar amounts of RNA (Figure 2)• Easy to use-just add the dye to sample, wait 5 minutes, and read• Suitable for use in 96- and 384-well plate formats• Compatible with most fluorescence-based microplate readers and fluorometers and FluorescenceApplications: The PicoGreen dsDNA Quantitation Reagent and Kits are ideal for PCR-based assays, microarray samples, DNA damage assays, enzyme activity assays, genomic DNA quantitation, measuring dsDNA in complex mixtures, and viral DNA quantitation.
    https://www.bioz.com/result/quant it picogreen assay kit/product/Thermo Fisher
    Average 93 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    quant it picogreen assay kit - by Bioz Stars, 2019-10
    93/100 stars

    Images

    1) Product Images from "Osteogenic Responses to Zirconia with Hydroxyapatite Coating by Aerosol Deposition"

    Article Title: Osteogenic Responses to Zirconia with Hydroxyapatite Coating by Aerosol Deposition

    Journal:

    doi: 10.1177/0022034514566432

    Cellular proliferation and viability on the discs. ( A ) Picogreen assay. Proliferation of MC3T3-E1 cells on titanium (Ti) or zirconia (Zr) discs on days 1, 4, and 7. ( B ) CCK-8 assay. Cell viability was tested on days 1, 4, and 7 under the same conditions
    Figure Legend Snippet: Cellular proliferation and viability on the discs. ( A ) Picogreen assay. Proliferation of MC3T3-E1 cells on titanium (Ti) or zirconia (Zr) discs on days 1, 4, and 7. ( B ) CCK-8 assay. Cell viability was tested on days 1, 4, and 7 under the same conditions

    Techniques Used: Picogreen Assay, CCK-8 Assay

    2) Product Images from "A Non-woven Path: Electrospun Poly(lactic acid) Scaffolds for Kidney Tissue Engineering"

    Article Title: A Non-woven Path: Electrospun Poly(lactic acid) Scaffolds for Kidney Tissue Engineering

    Journal: Tissue Engineering and Regenerative Medicine

    doi: 10.1007/s13770-017-0107-5

    DNA quantity per scaffold at 3 and 7 days, assessed by PicoGreen assay. This confirms the ability of all scaffold architectures to support primary kidney cell life. Analysis using a one-way ANOVA showed significant differences F(7,23) = 4.79, p = 0.002, post hoc Tukey analysis shows that was in regards to cryogenic scaffolds. Data presented as mean ± 95% confidence intervals, circles denote individual data points
    Figure Legend Snippet: DNA quantity per scaffold at 3 and 7 days, assessed by PicoGreen assay. This confirms the ability of all scaffold architectures to support primary kidney cell life. Analysis using a one-way ANOVA showed significant differences F(7,23) = 4.79, p = 0.002, post hoc Tukey analysis shows that was in regards to cryogenic scaffolds. Data presented as mean ± 95% confidence intervals, circles denote individual data points

    Techniques Used: Picogreen Assay

    Related Articles

    Centrifugation:

    Article Title: Preventive Effects of Carnosine on Lipopolysaccharide-induced Lung Injury
    Article Snippet: After centrifugation with a Cytospin® 4 (Thermo Fisher Scientific, Waltham, MA), cells were stained with Diff-Quik reagents and the ratio of neutrophils to total cell number was determined. .. The amount of protein and double-stranded DNA (dsDNA) present in the BALF was evaluated by the Bradford method and by using a Quant-iT™ PicoGreen® dsDNA Assay Kit (Thermo Fisher Scientific).

    Article Title: Nerve-Specific, Xenogeneic Extracellular Matrix Hydrogel Promotes Recovery Following Peripheral Nerve Injury
    Article Snippet: Protein content was removed from the sample by repeated phenol/chloroform/isoamyl alcohol (Thermo Scientific) extraction and centrifugation (10,000 G) until no protein precipitate was observed at the interface. .. Double-stranded DNA was then quantified using Quant-iT™ PicoGreen dsDNA assay kit (Life Technologies) following kit instructions.

    Picogreen Assay:

    Article Title: Exogenous DNA Loading into Extracellular Vesicles via Electroporation is Size-Dependent and Enables Limited Gene Delivery
    Article Snippet: Finally, EVs were resuspended in 100 µL of 1× TE by pipetting up and down several times. .. DNA quantification was performed using Quant-it PicoGreen Assay kit (Life Technologies, catalog #P7589) following the manufacturer’s protocol. .. Known quantities of DNA were labeled in the same fashion to establish a standard curve for quantification.

    Article Title: DNA polymerase-α regulates type I interferon activation through cytosolic RNA:DNA synthesis
    Article Snippet: The RNA:DNA samples were prepared as described above. .. The samples were quantified using Quant-iT™ PicoGreen® dsDNA Assay Kit (ThermoFisher Scientific, Grand Island, NY) according to manufacturer’s protocol. .. The samples were analyzed on 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA) using High Sensitivity DNA Analysis or RNA Analysis kits, and further quantified against internal standards as per the instruction manual.

    Article Title: Neutrophil extracellular traps in the host defense against sepsis induced by Burkholderia pseudomallei (melioidosis)
    Article Snippet: Since NET formation is quantified by the measurement of DNA release after DNase treatment, and DNA will degrade the epitope of the anti-nucleosome antibody, measurement of nucleosomes by this ELISA is not suitable [ ]. .. Therefore extracellular DNA was quantified in the supernatant using a Quant-iT™ Picogreen® dsDNA Assay Kit (Invitrogen) according to the manufacturer's instructions and was analyzed with a SpectraFluor Plus absorbance reader (Tecan Group, Männedorf, Switzerland). .. Mouse experiments were approved by the Academic Medical Center Animal Use and Welfare Committee (DIX16AB).

    Article Title: A Non-woven Path: Electrospun Poly(lactic acid) Scaffolds for Kidney Tissue Engineering
    Article Snippet: Scaffolds were washed 3 times in PBS and freeze dried for 24 h. Each scaffold was incubated within an ultrasonic bath (6 litre Cavitek Digital, The Allendale Group, UK) in 300 µl of a 2.5 units/ml papain digest solution, 5 mM cysteine HCL and 5 mM EDTA in PBS (all reagents from Sigma Aldrich, UK) at 60 °C for 24 h with periodic sonication. .. Total DNA content of the samples was calculated using a Quant-iT™ PicoGreen® assay kit (ThermoFisher, UK) as per the manufacturers’ instructions. .. Fluorescence was read using a microplate reader at Ex 490 nm Em 510–570 nm, N = 4 independent replicates.

    Article Title: Preventive Effects of Carnosine on Lipopolysaccharide-induced Lung Injury
    Article Snippet: After centrifugation with a Cytospin® 4 (Thermo Fisher Scientific, Waltham, MA), cells were stained with Diff-Quik reagents and the ratio of neutrophils to total cell number was determined. .. The amount of protein and double-stranded DNA (dsDNA) present in the BALF was evaluated by the Bradford method and by using a Quant-iT™ PicoGreen® dsDNA Assay Kit (Thermo Fisher Scientific). .. In vivo imaging of ROS in mice was performed as described previously , with some modifications.

    Article Title: Oxidized mitochondrial nucleoids released by neutrophils drive type I interferon production in human lupus
    Article Snippet: Results were expressed as mtDNA copy number ( ). .. When the DNA concentration was too low to perform ethanol precipitation/Real-Time PCR, the amount of extruded mtDNA was measured in the PK-digested supernatants with Quant-iT Picogreen dsDNA Assay kit (Invitrogen) as previously described ( ). .. Total DNA was extracted from neutrophils or monocytes with the DNAzol Reagent (Invitrogen).

    Article Title: Oxidized mitochondrial nucleoids released by neutrophils drive type I interferon production in human lupus
    Article Snippet: The DNA pellet was washed twice with 75% Ethanol, vacuum-dried, and then resuspended in 40 µl TE Buffer, pH 8. .. DNA concentration was assessed with Quant-iT Picogreen dsDNA Assay kit (Invitrogen). .. PCR was performed with 5 ng of isolated DNA, AmpliTaq Gold 360 (Invitrogen) and 0.5 µM of the following primers: mtDNA encoded NADH dehydrogenase subunit 1 (ND1; 5′-GCATTCCTAATGCTTACCGAAC-3′ and 5′-AAGGGTGGAGAGGTTAAAGGAG-3′); genomic DNA encoded Glyceraldehyde 3- phosphate dehydrogenase (GAPDH; 5′-AGGCAACTAGGATGGTGTGG-3′ and 5′-TTGATTTTGGAGGGATCTCG-3′).

    Article Title: Matrix-bound nanovesicles within ECM bioscaffolds
    Article Snippet: Collagenase from Clostridium histolyticum was obtained from Sigma-Aldrich. .. The proteinase K solution, Quant-iT PicoGreen dsDNA Assay Kit, and RNase A were obtained from Thermo Fisher Scientific. .. RNase-free DNase was obtained from Qiagen.

    Article Title: Nerve-Specific, Xenogeneic Extracellular Matrix Hydrogel Promotes Recovery Following Peripheral Nerve Injury
    Article Snippet: The solution was frozen using dried ice and centrifuged to produce a DNA pellet. .. Double-stranded DNA was then quantified using Quant-iT™ PicoGreen dsDNA assay kit (Life Technologies) following kit instructions. .. Statistical significance was assessed using one-way ANOVA with Tukey post-hoc test.

    Article Title: Perfusion decellularization of a human limb: A novel platform for composite tissue engineering and reconstructive surgery
    Article Snippet: Nuclease treatment was performed by treating native and acellular biopsies with Pierce Universal Nuclease (Thermo Fisher: 88701) at a concentration of 25 U / ml for 24 hours at room temperature. .. DNA extraction was performed using the Qiagen DNeasy blood and tissue kit and the double strand DNA concentration was assessed with the Quant-iT PicoGreen dsDNA assay (Life Technologies; P7589) following the manufacturer protocols. .. Fluorescence was read at 480nm/520nm (excitation/emission) and concentrations were calculated in relation to the standard curve.

    Article Title: Hydrogels derived from demineralized and decellularized bone extracellular matrix
    Article Snippet: The pellet was washed with ethanol, dried at room temperature and resuspended in 1 ml of TE buffer. .. The concentration of each extracted DNA sample was determined using a Quant-iT™ PicoGreen dsDNA assay kit (Invitrogen, Paisley, UK) following the manufacturer’s protocol. .. A standard curve was constructed by preparing samples of known DNA concentration from 0 to 1000 ng ml− 1 .

    Article Title: Lentiviral Vector Purification Using Genetically Encoded Biotin Mimic in Packaging Cell
    Article Snippet: After two rounds of cell division, recovered populations were assessed for sorting efficiency by flow cytometry staining using a BD LSRFORTESSA X-20 cell analyzer. .. Double-stranded DNA contamination was assessed in purified vectors using the Quant-iT PicoGreen dsDNA Assay Kit (Invitrogen). .. The manufacturers’ protocol was followed using a Thermo Scientific Nunc F96 MicroWell black polystyrene plate.

    Article Title: Blended electrospinning with human liver extracellular matrix for engineering new hepatic microenvironments
    Article Snippet: For each condition group, minimum n = 5. .. The Quant-IT Picogreen® dsDNA assay kit (Life Technologies™ ) was performed according to manufacturer instructions to establish the efficiency of the decellularization method and to estimate cell adherence and growth on the cell/scaffold constructs. .. Fluorescent intensity measurements were read in a Modulus™ II microplate reader (minimum n = 5) at an excitation wavelength of 480 nm and emission wavelength of 510–570 nm.

    Article Title: Growth factor and ultrasound-assisted bioreactor synergism for human mesenchymal stem cell chondrogenesis
    Article Snippet: Upon completion of the incubation, scaffolds were spun down, and supernatants were collected and subjected to DNA, GAG, and Hydroxyproline content measurements. .. Double-stranded DNA (dsDNA) was measured using Quant-iT™ PicoGreen® dsDNA Assay Kit (Life Technologies) according to manufacturer’s instructions. .. All quantifications were based on a λ DNA standard.

    Article Title: Posttranscriptional Upregulation of IDH1 by HuR Establishes a Powerful Survival Phenotype in Pancreatic Cancer Cells
    Article Snippet: Drugs were dissolved in DMSO, PBS, and cell culture media, respectively. .. Cells were plated in 96-well plates at 103 cells per well and assayed using Quant-it PicoGreen dsDNA Assay Kit (Invitrogen) at 5 days ( ). .. To estimate cell death, cells were trypsinized at 24 hours and counted after Trypan blue staining (Invitrogen) with a Hausser bright-line hemocytometer (Fisher Scientific).

    Article Title: Blended electrospinning with human liver extracellular matrix for engineering new hepatic microenvironments
    Article Snippet: The tissue was then washed with MilliQ H2 O with 0.1% 100U/ml penicillin, 100 µg/ml streptomycin, 0.25 µg/ml Fungizone® (amphotericin B) Anti-Anti solution (Gibco) for 4 hours and stored in sterile containers at −20 °C until use. .. Decellularization was confirmed using the Quant-IT™ Picogreen® dsDNA assay kit (Life Technologies™ ), performed according to manufacturer’s instructions. .. The ECM was lyophilized in a FreeZone® 4.5 freeze-drier (Labconco® ) before milling in a PM100® planetary ball mill (Retsch® ).

    Article Title: The Effects of Processing Methods upon Mechanical and Biologic Properties of Porcine Dermal Extracellular Matrix Scaffolds
    Article Snippet: Both decellularization methods were effective at decellularization of the dermis. .. Dermis treated with both the trypsin/SDS/TritonX-100 protocol (D-TST) and the trypsin/TritonX-100 protocol (d-TT) was determined to be effectively decellularized based upon the established criteria described earlier; specifically, (1) no nuclei observed by imaging and analysis of both DAPI and Hematoxylin and Eosin stained sections, (2) no DNA ≥200 base pairs observed by agarose gel analysis, and (3) samples had a content of < 50 ng dsDNA per mg initial dry weight as measured with the Quant-iT PicoGreen dsDNA Assay Kit (Invitrogen Corporation, Carlsbad, CA) ( ). .. The manufacture of a medical device from a biologic source presents unique challenges: contamination must be prevented, biologic variability must be controlled or at least understood, and a number of practical concerns such as sourcing, transportation of the raw material to a manufacturing plant, and preservation of tissue integrity must be addressed.

    Lambda DNA Preparation:

    Article Title: Nonviral Gene Delivery of Growth and Differentiation Factor 5 to Human Mesenchymal Stem Cells Injected into a 3D Bovine Intervertebral Disc Organ Culture System
    Article Snippet: Standard curve is done with a dilution of chondroitin sulfate (C9819, Sigma-Aldrich) in papain buffer, described above. .. The amount of DNA was determined by Quant-iT PicoGreen dsDNA Reagent (P11496, Invitrogen, Life Technologies Corp., Basel, Switzerland), following the manufacturer's protocol with a high-range standard curve of Lambda DNA standard (P11496, Invitrogen Life Technologies Corp., Basel, Switzerland) [ , ]. .. Fluorescence was measured using a spectrofluorometer reader (SpectraMax, M5, Molecular Devices, USA) at excitation 487 nm and emission at 525 nm, with a cutoff at 515 nm.

    Construct:

    Article Title: Blended electrospinning with human liver extracellular matrix for engineering new hepatic microenvironments
    Article Snippet: For each condition group, minimum n = 5. .. The Quant-IT Picogreen® dsDNA assay kit (Life Technologies™ ) was performed according to manufacturer instructions to establish the efficiency of the decellularization method and to estimate cell adherence and growth on the cell/scaffold constructs. .. Fluorescent intensity measurements were read in a Modulus™ II microplate reader (minimum n = 5) at an excitation wavelength of 480 nm and emission wavelength of 510–570 nm.

    Real-time Polymerase Chain Reaction:

    Article Title: Oxidized mitochondrial nucleoids released by neutrophils drive type I interferon production in human lupus
    Article Snippet: 3 ng of DNA isolated from cell-free supernatants were subjected to Real-Time PCR with Power SYBR Green PCR Master Mix (Invitrogen) and 0.5 µM of the primers described above. .. When the DNA concentration was too low to perform ethanol precipitation/Real-Time PCR, the amount of extruded mtDNA was measured in the PK-digested supernatants with Quant-iT Picogreen dsDNA Assay kit (Invitrogen) as previously described ( ).

    Incubation:

    Article Title: Functional characterization of an alkaline exonuclease and single strand annealing protein from the SXT genetic element of Vibrio cholerae
    Article Snippet: Aliquots (20 μl) were quenched (1% SDS + 20 mM EDTA) at the times indicated over a 160 min period, then analyzed on 1% agarose TAE gels. .. The double strand exonuclease activities of SXT-Exo and lambda-Exo under various conditions were determined by quantifying the amounts of double strand DNA that remained after enzymatic incubation, using the PicoGreen DNA fluorescence reagent (Invitrogen, catalogue # P7589). .. These quenched PicoGreen fluorescence assays were performed as described previously [ , ] with minor modifications.

    Article Title: Exogenous DNA Loading into Extracellular Vesicles via Electroporation is Size-Dependent and Enables Limited Gene Delivery
    Article Snippet: Samples were then incubated at room temperature for 15 min and subsequently transferred into Nanosep tubes and centrifuged at 5000 g at 4 °C for 5 min to remove buffer and unincorporated dsDNA. .. DNA quantification was performed using Quant-it PicoGreen Assay kit (Life Technologies, catalog #P7589) following the manufacturer’s protocol.

    Article Title: A Non-woven Path: Electrospun Poly(lactic acid) Scaffolds for Kidney Tissue Engineering
    Article Snippet: Scaffolds were washed 3 times in PBS and freeze dried for 24 h. Each scaffold was incubated within an ultrasonic bath (6 litre Cavitek Digital, The Allendale Group, UK) in 300 µl of a 2.5 units/ml papain digest solution, 5 mM cysteine HCL and 5 mM EDTA in PBS (all reagents from Sigma Aldrich, UK) at 60 °C for 24 h with periodic sonication. .. Total DNA content of the samples was calculated using a Quant-iT™ PicoGreen® assay kit (ThermoFisher, UK) as per the manufacturers’ instructions.

    Article Title: Oxidized mitochondrial nucleoids released by neutrophils drive type I interferon production in human lupus
    Article Snippet: DNA concentration was assessed with Quant-iT Picogreen dsDNA Assay kit (Invitrogen). .. DNA concentration was assessed with Quant-iT Picogreen dsDNA Assay kit (Invitrogen).

    Article Title: Perfusion decellularization of a human limb: A novel platform for composite tissue engineering and reconstructive surgery
    Article Snippet: DNA extraction was performed using the Qiagen DNeasy blood and tissue kit and the double strand DNA concentration was assessed with the Quant-iT PicoGreen dsDNA assay (Life Technologies; P7589) following the manufacturer protocols. .. DNA extraction was performed using the Qiagen DNeasy blood and tissue kit and the double strand DNA concentration was assessed with the Quant-iT PicoGreen dsDNA assay (Life Technologies; P7589) following the manufacturer protocols.

    Article Title: Growth factor and ultrasound-assisted bioreactor synergism for human mesenchymal stem cell chondrogenesis
    Article Snippet: Upon completion of the incubation, scaffolds were spun down, and supernatants were collected and subjected to DNA, GAG, and Hydroxyproline content measurements. .. Double-stranded DNA (dsDNA) was measured using Quant-iT™ PicoGreen® dsDNA Assay Kit (Life Technologies) according to manufacturer’s instructions.

    Activity Assay:

    Article Title: Exogenous DNA Loading into Extracellular Vesicles via Electroporation is Size-Dependent and Enables Limited Gene Delivery
    Article Snippet: To remove excess unbound dsDNA, samples were digested in the same tube by adding DNase I enzyme (1.5 Kunitz units/µL, Qiagen #79254) and incubating for 20 min. DNase I activity was confirmed by digesting an equivalent amount of loaded DNA in solution. .. DNA quantification was performed using Quant-it PicoGreen Assay kit (Life Technologies, catalog #P7589) following the manufacturer’s protocol.

    Electroporation:

    Article Title: Exogenous DNA Loading into Extracellular Vesicles via Electroporation is Size-Dependent and Enables Limited Gene Delivery
    Article Snippet: Paragraph title: DNA Loading into EVs by Electroporation ... DNA quantification was performed using Quant-it PicoGreen Assay kit (Life Technologies, catalog #P7589) following the manufacturer’s protocol.

    SYBR Green Assay:

    Article Title: Oxidized mitochondrial nucleoids released by neutrophils drive type I interferon production in human lupus
    Article Snippet: 3 ng of DNA isolated from cell-free supernatants were subjected to Real-Time PCR with Power SYBR Green PCR Master Mix (Invitrogen) and 0.5 µM of the primers described above. .. When the DNA concentration was too low to perform ethanol precipitation/Real-Time PCR, the amount of extruded mtDNA was measured in the PK-digested supernatants with Quant-iT Picogreen dsDNA Assay kit (Invitrogen) as previously described ( ).

    Cell Culture:

    Article Title: Posttranscriptional Upregulation of IDH1 by HuR Establishes a Powerful Survival Phenotype in Pancreatic Cancer Cells
    Article Snippet: Drugs were dissolved in DMSO, PBS, and cell culture media, respectively. .. Cells were plated in 96-well plates at 103 cells per well and assayed using Quant-it PicoGreen dsDNA Assay Kit (Invitrogen) at 5 days ( ).

    Collagen Assay:

    Article Title: Perfusion decellularization of a human limb: A novel platform for composite tissue engineering and reconstructive surgery
    Article Snippet: DNA extraction was performed using the Qiagen DNeasy blood and tissue kit and the double strand DNA concentration was assessed with the Quant-iT PicoGreen dsDNA assay (Life Technologies; P7589) following the manufacturer protocols. .. DNA extraction was performed using the Qiagen DNeasy blood and tissue kit and the double strand DNA concentration was assessed with the Quant-iT PicoGreen dsDNA assay (Life Technologies; P7589) following the manufacturer protocols.

    Imaging:

    Article Title: The Effects of Processing Methods upon Mechanical and Biologic Properties of Porcine Dermal Extracellular Matrix Scaffolds
    Article Snippet: Both decellularization methods were effective at decellularization of the dermis. .. Dermis treated with both the trypsin/SDS/TritonX-100 protocol (D-TST) and the trypsin/TritonX-100 protocol (d-TT) was determined to be effectively decellularized based upon the established criteria described earlier; specifically, (1) no nuclei observed by imaging and analysis of both DAPI and Hematoxylin and Eosin stained sections, (2) no DNA ≥200 base pairs observed by agarose gel analysis, and (3) samples had a content of < 50 ng dsDNA per mg initial dry weight as measured with the Quant-iT PicoGreen dsDNA Assay Kit (Invitrogen Corporation, Carlsbad, CA) ( ). .. The manufacture of a medical device from a biologic source presents unique challenges: contamination must be prevented, biologic variability must be controlled or at least understood, and a number of practical concerns such as sourcing, transportation of the raw material to a manufacturing plant, and preservation of tissue integrity must be addressed.

    Polymerase Chain Reaction:

    Article Title: Oxidized mitochondrial nucleoids released by neutrophils drive type I interferon production in human lupus
    Article Snippet: Results were expressed as mtDNA copy number ( ). .. When the DNA concentration was too low to perform ethanol precipitation/Real-Time PCR, the amount of extruded mtDNA was measured in the PK-digested supernatants with Quant-iT Picogreen dsDNA Assay kit (Invitrogen) as previously described ( ). .. Total DNA was extracted from neutrophils or monocytes with the DNAzol Reagent (Invitrogen).

    Article Title: Oxidized mitochondrial nucleoids released by neutrophils drive type I interferon production in human lupus
    Article Snippet: DNA concentration was assessed with Quant-iT Picogreen dsDNA Assay kit (Invitrogen). .. PCR was performed with 5 ng of isolated DNA, AmpliTaq Gold 360 (Invitrogen) and 0.5 µM of the following primers: mtDNA encoded NADH dehydrogenase subunit 1 (ND1; 5′-GCATTCCTAATGCTTACCGAAC-3′ and 5′-AAGGGTGGAGAGGTTAAAGGAG-3′); genomic DNA encoded Glyceraldehyde 3- phosphate dehydrogenase (GAPDH; 5′-AGGCAACTAGGATGGTGTGG-3′ and 5′-TTGATTTTGGAGGGATCTCG-3′).

    Sonication:

    Article Title: A Non-woven Path: Electrospun Poly(lactic acid) Scaffolds for Kidney Tissue Engineering
    Article Snippet: Scaffolds were washed 3 times in PBS and freeze dried for 24 h. Each scaffold was incubated within an ultrasonic bath (6 litre Cavitek Digital, The Allendale Group, UK) in 300 µl of a 2.5 units/ml papain digest solution, 5 mM cysteine HCL and 5 mM EDTA in PBS (all reagents from Sigma Aldrich, UK) at 60 °C for 24 h with periodic sonication. .. Total DNA content of the samples was calculated using a Quant-iT™ PicoGreen® assay kit (ThermoFisher, UK) as per the manufacturers’ instructions.

    Binding Assay:

    Article Title: Nonviral Gene Delivery of Growth and Differentiation Factor 5 to Human Mesenchymal Stem Cells Injected into a 3D Bovine Intervertebral Disc Organ Culture System
    Article Snippet: Glycosaminoglycan (GAG) content was quantified by 1,9-dimethylmethylene blue (DMMB) binding assay. .. The amount of DNA was determined by Quant-iT PicoGreen dsDNA Reagent (P11496, Invitrogen, Life Technologies Corp., Basel, Switzerland), following the manufacturer's protocol with a high-range standard curve of Lambda DNA standard (P11496, Invitrogen Life Technologies Corp., Basel, Switzerland) [ , ].

    Molecular Weight:

    Article Title: Exogenous DNA Loading into Extracellular Vesicles via Electroporation is Size-Dependent and Enables Limited Gene Delivery
    Article Snippet: DNA quantification was performed using Quant-it PicoGreen Assay kit (Life Technologies, catalog #P7589) following the manufacturer’s protocol. .. Known quantities of DNA were labeled in the same fashion to establish a standard curve for quantification.

    DNA Extraction:

    Article Title: Perfusion decellularization of a human limb: A novel platform for composite tissue engineering and reconstructive surgery
    Article Snippet: Nuclease treatment was performed by treating native and acellular biopsies with Pierce Universal Nuclease (Thermo Fisher: 88701) at a concentration of 25 U / ml for 24 hours at room temperature. .. DNA extraction was performed using the Qiagen DNeasy blood and tissue kit and the double strand DNA concentration was assessed with the Quant-iT PicoGreen dsDNA assay (Life Technologies; P7589) following the manufacturer protocols. .. Fluorescence was read at 480nm/520nm (excitation/emission) and concentrations were calculated in relation to the standard curve.

    Fluorescence:

    Article Title: Functional characterization of an alkaline exonuclease and single strand annealing protein from the SXT genetic element of Vibrio cholerae
    Article Snippet: Aliquots (20 μl) were quenched (1% SDS + 20 mM EDTA) at the times indicated over a 160 min period, then analyzed on 1% agarose TAE gels. .. The double strand exonuclease activities of SXT-Exo and lambda-Exo under various conditions were determined by quantifying the amounts of double strand DNA that remained after enzymatic incubation, using the PicoGreen DNA fluorescence reagent (Invitrogen, catalogue # P7589). .. These quenched PicoGreen fluorescence assays were performed as described previously [ , ] with minor modifications.

    Article Title: Blended electrospinning with human liver extracellular matrix for engineering new hepatic microenvironments
    Article Snippet: The Quant-IT Picogreen® dsDNA assay kit (Life Technologies™ ) was performed according to manufacturer instructions to establish the efficiency of the decellularization method and to estimate cell adherence and growth on the cell/scaffold constructs. .. The Quant-IT Picogreen® dsDNA assay kit (Life Technologies™ ) was performed according to manufacturer instructions to establish the efficiency of the decellularization method and to estimate cell adherence and growth on the cell/scaffold constructs.

    Isolation:

    Article Title: Oxidized mitochondrial nucleoids released by neutrophils drive type I interferon production in human lupus
    Article Snippet: 3 ng of DNA isolated from cell-free supernatants were subjected to Real-Time PCR with Power SYBR Green PCR Master Mix (Invitrogen) and 0.5 µM of the primers described above. .. When the DNA concentration was too low to perform ethanol precipitation/Real-Time PCR, the amount of extruded mtDNA was measured in the PK-digested supernatants with Quant-iT Picogreen dsDNA Assay kit (Invitrogen) as previously described ( ).

    Article Title: Oxidized mitochondrial nucleoids released by neutrophils drive type I interferon production in human lupus
    Article Snippet: Paragraph title: Isolation of DNA from supernatants and analysis ... DNA concentration was assessed with Quant-iT Picogreen dsDNA Assay kit (Invitrogen).

    Microscopy:

    Article Title: Neutrophil extracellular traps in the host defense against sepsis induced by Burkholderia pseudomallei (melioidosis)
    Article Snippet: NETs were stained with Sytox Green 5 mM (Molecular Probes, Eugene, OR, USA) and then visualized using a confocal microscope (Olympus IX81, Olympus Corporation, Tokyo, Japan). .. Therefore extracellular DNA was quantified in the supernatant using a Quant-iT™ Picogreen® dsDNA Assay Kit (Invitrogen) according to the manufacturer's instructions and was analyzed with a SpectraFluor Plus absorbance reader (Tecan Group, Männedorf, Switzerland).

    Purification:

    Article Title: Lentiviral Vector Purification Using Genetically Encoded Biotin Mimic in Packaging Cell
    Article Snippet: After two rounds of cell division, recovered populations were assessed for sorting efficiency by flow cytometry staining using a BD LSRFORTESSA X-20 cell analyzer. .. Double-stranded DNA contamination was assessed in purified vectors using the Quant-iT PicoGreen dsDNA Assay Kit (Invitrogen). .. The manufacturers’ protocol was followed using a Thermo Scientific Nunc F96 MicroWell black polystyrene plate.

    Dot Blot:

    Article Title: Oxidized mitochondrial nucleoids released by neutrophils drive type I interferon production in human lupus
    Article Snippet: DNA concentration was assessed with Quant-iT Picogreen dsDNA Assay kit (Invitrogen). .. DNA concentration was assessed with Quant-iT Picogreen dsDNA Assay kit (Invitrogen).

    Diff-Quik:

    Article Title: Preventive Effects of Carnosine on Lipopolysaccharide-induced Lung Injury
    Article Snippet: After centrifugation with a Cytospin® 4 (Thermo Fisher Scientific, Waltham, MA), cells were stained with Diff-Quik reagents and the ratio of neutrophils to total cell number was determined. .. The amount of protein and double-stranded DNA (dsDNA) present in the BALF was evaluated by the Bradford method and by using a Quant-iT™ PicoGreen® dsDNA Assay Kit (Thermo Fisher Scientific).

    Software:

    Article Title: Lentiviral Vector Purification Using Genetically Encoded Biotin Mimic in Packaging Cell
    Article Snippet: Double-stranded DNA contamination was assessed in purified vectors using the Quant-iT PicoGreen dsDNA Assay Kit (Invitrogen). .. Double-stranded DNA contamination was assessed in purified vectors using the Quant-iT PicoGreen dsDNA Assay Kit (Invitrogen).

    Microplate Reader Absorbance Measurement:

    Article Title: Neutrophil extracellular traps in the host defense against sepsis induced by Burkholderia pseudomallei (melioidosis)
    Article Snippet: Since NET formation is quantified by the measurement of DNA release after DNase treatment, and DNA will degrade the epitope of the anti-nucleosome antibody, measurement of nucleosomes by this ELISA is not suitable [ ]. .. Therefore extracellular DNA was quantified in the supernatant using a Quant-iT™ Picogreen® dsDNA Assay Kit (Invitrogen) according to the manufacturer's instructions and was analyzed with a SpectraFluor Plus absorbance reader (Tecan Group, Männedorf, Switzerland). .. Mouse experiments were approved by the Academic Medical Center Animal Use and Welfare Committee (DIX16AB).

    Hydroxyproline Assay:

    Article Title: Growth factor and ultrasound-assisted bioreactor synergism for human mesenchymal stem cell chondrogenesis
    Article Snippet: Double-stranded DNA (dsDNA) was measured using Quant-iT™ PicoGreen® dsDNA Assay Kit (Life Technologies) according to manufacturer’s instructions. .. Total GAG content was determined with DMMB assay on papain-digested supernatant according to the protocol described elsewhere and quantified based on shark chondroitin sulfate standard.

    Irradiation:

    Article Title: Oxidized mitochondrial nucleoids released by neutrophils drive type I interferon production in human lupus
    Article Snippet: DNA concentration was assessed with Quant-iT Picogreen dsDNA Assay kit (Invitrogen). .. DNA concentration was assessed with Quant-iT Picogreen dsDNA Assay kit (Invitrogen).

    Agarose Gel Electrophoresis:

    Article Title: Oxidized mitochondrial nucleoids released by neutrophils drive type I interferon production in human lupus
    Article Snippet: DNA concentration was assessed with Quant-iT Picogreen dsDNA Assay kit (Invitrogen). .. PCR was performed with 5 ng of isolated DNA, AmpliTaq Gold 360 (Invitrogen) and 0.5 µM of the following primers: mtDNA encoded NADH dehydrogenase subunit 1 (ND1; 5′-GCATTCCTAATGCTTACCGAAC-3′ and 5′-AAGGGTGGAGAGGTTAAAGGAG-3′); genomic DNA encoded Glyceraldehyde 3- phosphate dehydrogenase (GAPDH; 5′-AGGCAACTAGGATGGTGTGG-3′ and 5′-TTGATTTTGGAGGGATCTCG-3′).

    Article Title: Nerve-Specific, Xenogeneic Extracellular Matrix Hydrogel Promotes Recovery Following Peripheral Nerve Injury
    Article Snippet: Double-stranded DNA was then quantified using Quant-iT™ PicoGreen dsDNA assay kit (Life Technologies) following kit instructions. .. Double-stranded DNA was then quantified using Quant-iT™ PicoGreen dsDNA assay kit (Life Technologies) following kit instructions.

    Article Title: The Effects of Processing Methods upon Mechanical and Biologic Properties of Porcine Dermal Extracellular Matrix Scaffolds
    Article Snippet: Both decellularization methods were effective at decellularization of the dermis. .. Dermis treated with both the trypsin/SDS/TritonX-100 protocol (D-TST) and the trypsin/TritonX-100 protocol (d-TT) was determined to be effectively decellularized based upon the established criteria described earlier; specifically, (1) no nuclei observed by imaging and analysis of both DAPI and Hematoxylin and Eosin stained sections, (2) no DNA ≥200 base pairs observed by agarose gel analysis, and (3) samples had a content of < 50 ng dsDNA per mg initial dry weight as measured with the Quant-iT PicoGreen dsDNA Assay Kit (Invitrogen Corporation, Carlsbad, CA) ( ). .. The manufacture of a medical device from a biologic source presents unique challenges: contamination must be prevented, biologic variability must be controlled or at least understood, and a number of practical concerns such as sourcing, transportation of the raw material to a manufacturing plant, and preservation of tissue integrity must be addressed.

    Electrophoresis:

    Article Title: DNA polymerase-α regulates type I interferon activation through cytosolic RNA:DNA synthesis
    Article Snippet: Paragraph title: On-chip DNA and RNA capillary electrophoresis ... The samples were quantified using Quant-iT™ PicoGreen® dsDNA Assay Kit (ThermoFisher Scientific, Grand Island, NY) according to manufacturer’s protocol.

    Enzyme-linked Immunosorbent Assay:

    Article Title: Neutrophil extracellular traps in the host defense against sepsis induced by Burkholderia pseudomallei (melioidosis)
    Article Snippet: Since NET formation is quantified by the measurement of DNA release after DNase treatment, and DNA will degrade the epitope of the anti-nucleosome antibody, measurement of nucleosomes by this ELISA is not suitable [ ]. .. Therefore extracellular DNA was quantified in the supernatant using a Quant-iT™ Picogreen® dsDNA Assay Kit (Invitrogen) according to the manufacturer's instructions and was analyzed with a SpectraFluor Plus absorbance reader (Tecan Group, Männedorf, Switzerland).

    Concentration Assay:

    Article Title: Exogenous DNA Loading into Extracellular Vesicles via Electroporation is Size-Dependent and Enables Limited Gene Delivery
    Article Snippet: After digestion, DNase I was inactivated by adding EDTA to a final concentration of 20 mM. .. DNA quantification was performed using Quant-it PicoGreen Assay kit (Life Technologies, catalog #P7589) following the manufacturer’s protocol.

    Article Title: Oxidized mitochondrial nucleoids released by neutrophils drive type I interferon production in human lupus
    Article Snippet: Results were expressed as mtDNA copy number ( ). .. When the DNA concentration was too low to perform ethanol precipitation/Real-Time PCR, the amount of extruded mtDNA was measured in the PK-digested supernatants with Quant-iT Picogreen dsDNA Assay kit (Invitrogen) as previously described ( ). .. Total DNA was extracted from neutrophils or monocytes with the DNAzol Reagent (Invitrogen).

    Article Title: Oxidized mitochondrial nucleoids released by neutrophils drive type I interferon production in human lupus
    Article Snippet: The DNA pellet was washed twice with 75% Ethanol, vacuum-dried, and then resuspended in 40 µl TE Buffer, pH 8. .. DNA concentration was assessed with Quant-iT Picogreen dsDNA Assay kit (Invitrogen). .. PCR was performed with 5 ng of isolated DNA, AmpliTaq Gold 360 (Invitrogen) and 0.5 µM of the following primers: mtDNA encoded NADH dehydrogenase subunit 1 (ND1; 5′-GCATTCCTAATGCTTACCGAAC-3′ and 5′-AAGGGTGGAGAGGTTAAAGGAG-3′); genomic DNA encoded Glyceraldehyde 3- phosphate dehydrogenase (GAPDH; 5′-AGGCAACTAGGATGGTGTGG-3′ and 5′-TTGATTTTGGAGGGATCTCG-3′).

    Article Title: Perfusion decellularization of a human limb: A novel platform for composite tissue engineering and reconstructive surgery
    Article Snippet: Nuclease treatment was performed by treating native and acellular biopsies with Pierce Universal Nuclease (Thermo Fisher: 88701) at a concentration of 25 U / ml for 24 hours at room temperature. .. DNA extraction was performed using the Qiagen DNeasy blood and tissue kit and the double strand DNA concentration was assessed with the Quant-iT PicoGreen dsDNA assay (Life Technologies; P7589) following the manufacturer protocols. .. Fluorescence was read at 480nm/520nm (excitation/emission) and concentrations were calculated in relation to the standard curve.

    Article Title: Hydrogels derived from demineralized and decellularized bone extracellular matrix
    Article Snippet: The pellet was washed with ethanol, dried at room temperature and resuspended in 1 ml of TE buffer. .. The concentration of each extracted DNA sample was determined using a Quant-iT™ PicoGreen dsDNA assay kit (Invitrogen, Paisley, UK) following the manufacturer’s protocol. .. A standard curve was constructed by preparing samples of known DNA concentration from 0 to 1000 ng ml− 1 .

    Article Title: Blended electrospinning with human liver extracellular matrix for engineering new hepatic microenvironments
    Article Snippet: The Quant-IT Picogreen® dsDNA assay kit (Life Technologies™ ) was performed according to manufacturer instructions to establish the efficiency of the decellularization method and to estimate cell adherence and growth on the cell/scaffold constructs. .. The Quant-IT Picogreen® dsDNA assay kit (Life Technologies™ ) was performed according to manufacturer instructions to establish the efficiency of the decellularization method and to estimate cell adherence and growth on the cell/scaffold constructs.

    Transmission Electron Microscopy:

    Article Title: Matrix-bound nanovesicles within ECM bioscaffolds
    Article Snippet: The proteinase K solution, Quant-iT PicoGreen dsDNA Assay Kit, and RNase A were obtained from Thermo Fisher Scientific. .. The proteinase K solution, Quant-iT PicoGreen dsDNA Assay Kit, and RNase A were obtained from Thermo Fisher Scientific.

    Standard Deviation:

    Article Title: Functional characterization of an alkaline exonuclease and single strand annealing protein from the SXT genetic element of Vibrio cholerae
    Article Snippet: The double strand exonuclease activities of SXT-Exo and lambda-Exo under various conditions were determined by quantifying the amounts of double strand DNA that remained after enzymatic incubation, using the PicoGreen DNA fluorescence reagent (Invitrogen, catalogue # P7589). .. The double strand exonuclease activities of SXT-Exo and lambda-Exo under various conditions were determined by quantifying the amounts of double strand DNA that remained after enzymatic incubation, using the PicoGreen DNA fluorescence reagent (Invitrogen, catalogue # P7589).

    Staining:

    Article Title: Neutrophil extracellular traps in the host defense against sepsis induced by Burkholderia pseudomallei (melioidosis)
    Article Snippet: NETs were stained with Sytox Green 5 mM (Molecular Probes, Eugene, OR, USA) and then visualized using a confocal microscope (Olympus IX81, Olympus Corporation, Tokyo, Japan). .. Therefore extracellular DNA was quantified in the supernatant using a Quant-iT™ Picogreen® dsDNA Assay Kit (Invitrogen) according to the manufacturer's instructions and was analyzed with a SpectraFluor Plus absorbance reader (Tecan Group, Männedorf, Switzerland).

    Article Title: Preventive Effects of Carnosine on Lipopolysaccharide-induced Lung Injury
    Article Snippet: After centrifugation with a Cytospin® 4 (Thermo Fisher Scientific, Waltham, MA), cells were stained with Diff-Quik reagents and the ratio of neutrophils to total cell number was determined. .. The amount of protein and double-stranded DNA (dsDNA) present in the BALF was evaluated by the Bradford method and by using a Quant-iT™ PicoGreen® dsDNA Assay Kit (Thermo Fisher Scientific).

    Article Title: Nerve-Specific, Xenogeneic Extracellular Matrix Hydrogel Promotes Recovery Following Peripheral Nerve Injury
    Article Snippet: Additional samples were stained using Luxol fast blue to determine removal of myelin as an additional metric of decellularization. .. Double-stranded DNA was then quantified using Quant-iT™ PicoGreen dsDNA assay kit (Life Technologies) following kit instructions.

    Article Title: The Effects of Processing Methods upon Mechanical and Biologic Properties of Porcine Dermal Extracellular Matrix Scaffolds
    Article Snippet: Both decellularization methods were effective at decellularization of the dermis. .. Dermis treated with both the trypsin/SDS/TritonX-100 protocol (D-TST) and the trypsin/TritonX-100 protocol (d-TT) was determined to be effectively decellularized based upon the established criteria described earlier; specifically, (1) no nuclei observed by imaging and analysis of both DAPI and Hematoxylin and Eosin stained sections, (2) no DNA ≥200 base pairs observed by agarose gel analysis, and (3) samples had a content of < 50 ng dsDNA per mg initial dry weight as measured with the Quant-iT PicoGreen dsDNA Assay Kit (Invitrogen Corporation, Carlsbad, CA) ( ). .. The manufacture of a medical device from a biologic source presents unique challenges: contamination must be prevented, biologic variability must be controlled or at least understood, and a number of practical concerns such as sourcing, transportation of the raw material to a manufacturing plant, and preservation of tissue integrity must be addressed.

    Dimethylmethylene Blue Assay:

    Article Title: Growth factor and ultrasound-assisted bioreactor synergism for human mesenchymal stem cell chondrogenesis
    Article Snippet: Double-stranded DNA (dsDNA) was measured using Quant-iT™ PicoGreen® dsDNA Assay Kit (Life Technologies) according to manufacturer’s instructions. .. Double-stranded DNA (dsDNA) was measured using Quant-iT™ PicoGreen® dsDNA Assay Kit (Life Technologies) according to manufacturer’s instructions.

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  • 99
    Thermo Fisher picogreen
    Quantitative detection of DNA damage in C . neoformans H99 and S . cerevisiae BY4741 exposed to ionizing radiation using the long mitochondrial DNA and nuclear DNA fragments. H99 and BY4741 cells were exposed to ionizing radiation from doses of 500 Gy to 3000 Gy. Irradiated cells were immediately frozen at -80 ° C and DNA extraction was performed as described in the Material and Methods. PCR products were quantified using the <t>PicoGreen</t> assay. DNA lesions were calculated according to the formula (lesions/amplified fragment = -ln (A D /A C )). The solid lines are lines connecting each data point. The dotted lines are the linear regression lines. (A and D) long mitochondrial DNA fragment lesions, (B and E) long nuclear DNA fragment lesions, (C and F) short mitochondrial DNA fragment PCR yield. Results were from three independent experiments with triplicate samples in each experiment (3 samples/point/experiment x 3). Mean ± S.E.M. **, p
    Picogreen, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 213 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/picogreen/product/Thermo Fisher
    Average 99 stars, based on 213 article reviews
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    99
    Thermo Fisher quant it picogreen kit
    Murine and human neutrophils release more dsDNA when exposed to L. (V) p . strains that are resistant to either MA or MIL. Bone marrow derived murine neutrophils (A) and blood-derived human neutrophils from healthy donors (B) were exposed to L. (V.) p . of the indicated susceptibility at a 5:1 MOI. As controls, incubations with PMA, PMA + DNase or without stimulus were also performed. Four hours later, NET formation was quantified by measuring the levels of double stranded DNA (dsDNA) released in the supernatant using the <t>PicoGreen</t> fluorescent dye assay. * p
    Quant It Picogreen Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quant it picogreen kit/product/Thermo Fisher
    Average 99 stars, based on 18 article reviews
    Price from $9.99 to $1999.99
    quant it picogreen kit - by Bioz Stars, 2019-10
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    99
    Thermo Fisher quant it picogreen dsdna assay kit
    POLA1 deficiency is associated with reduced cytosolic RNA:DNA levels ( a ) Immunoprecipitation of <t>dsDNA</t> and RNA:DNA from cytosolic fractions of HEK293T cells using the corresponding antibodies and Protein-G agarose beads. Beads were stained with <t>Picogreen</t> ® and visualized by fluorescence microscopy. Nucleic acid recovery was quantified by image analysis (ImageJ). Mean relative fluorescence levels are indicated in parantheses. ( b ) As in (a), dsDNA or RNA:DNA were quantified in control or XLPDR-derived fibroblasts. ( c ) RNA:DNA was quantified in a similar manner in three models of POLA1 deficiency: HEK293T and HeLa after POLA1 silencing (compared to control siRNA), and XLPDR-derived dermal fibroblasts (compared to control cells). Representative images are shown and relative fluorescence levels are indicated. ( d ) Cytosolic RNA:DNA content was quantified in XLPDR-derived fibroblasts stably transduced with a lentivirus encoding HA-tagged wild-type POLA1 (HA-POLA1wt) or EV. Representative images and relative fluorescence levels are indicated. ( e ) Immunoblot analysis of total and p-TBK1, and total and HA-tagged POLA1 expression in XLPDR fibroblast stably expressing HA-POLA1wt or EV (as in d). ( f ) qRT-PCR analysis of IFIT1 mRNA expression in the same cells depicted in (d); when indicated, the cells were treated with poly(dA:dT) for 16h. Scale bars are 20 μm. *p≤0.05, **p≤0.01, NS – not significant (unpaired Student’s t -test). In ( f ) all comparisons are made against the corresponding WT conditions. Data are representative of 1 ( d,e ), 2 ( a,b,f ), or 4 ( c ) independent experiments (mean ( a,c,d ) and mean and s.e.m.( b,f )).
    Quant It Picogreen Dsdna Assay Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1235 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quant it picogreen dsdna assay kit/product/Thermo Fisher
    Average 99 stars, based on 1235 article reviews
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    quant it picogreen dsdna assay kit - by Bioz Stars, 2019-10
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    Quantitative detection of DNA damage in C . neoformans H99 and S . cerevisiae BY4741 exposed to ionizing radiation using the long mitochondrial DNA and nuclear DNA fragments. H99 and BY4741 cells were exposed to ionizing radiation from doses of 500 Gy to 3000 Gy. Irradiated cells were immediately frozen at -80 ° C and DNA extraction was performed as described in the Material and Methods. PCR products were quantified using the PicoGreen assay. DNA lesions were calculated according to the formula (lesions/amplified fragment = -ln (A D /A C )). The solid lines are lines connecting each data point. The dotted lines are the linear regression lines. (A and D) long mitochondrial DNA fragment lesions, (B and E) long nuclear DNA fragment lesions, (C and F) short mitochondrial DNA fragment PCR yield. Results were from three independent experiments with triplicate samples in each experiment (3 samples/point/experiment x 3). Mean ± S.E.M. **, p

    Journal: PLoS ONE

    Article Title: Measuring radiation-induced DNA damage in Cryptococcus neoformans and Saccharomyces cerevisiae using long range quantitative PCR

    doi: 10.1371/journal.pone.0207071

    Figure Lengend Snippet: Quantitative detection of DNA damage in C . neoformans H99 and S . cerevisiae BY4741 exposed to ionizing radiation using the long mitochondrial DNA and nuclear DNA fragments. H99 and BY4741 cells were exposed to ionizing radiation from doses of 500 Gy to 3000 Gy. Irradiated cells were immediately frozen at -80 ° C and DNA extraction was performed as described in the Material and Methods. PCR products were quantified using the PicoGreen assay. DNA lesions were calculated according to the formula (lesions/amplified fragment = -ln (A D /A C )). The solid lines are lines connecting each data point. The dotted lines are the linear regression lines. (A and D) long mitochondrial DNA fragment lesions, (B and E) long nuclear DNA fragment lesions, (C and F) short mitochondrial DNA fragment PCR yield. Results were from three independent experiments with triplicate samples in each experiment (3 samples/point/experiment x 3). Mean ± S.E.M. **, p

    Article Snippet: DNA sample concentrations were determined by fluorescence measurements after DNA staining with PicoGreen (Quant-iT PicoGreen dsDNA reagent and kit; Thermo Fisher Scientific, New York, NY).

    Techniques: Irradiation, DNA Extraction, Polymerase Chain Reaction, Picogreen Assay, Amplification

    Quantitative amplification of target DNA fragments in LR-QPCR with respect to the amount of template DNA. DNA isolated from C . neoformans H99 and S . cerevisiae BY4741 were serially diluted with nuclease-free water for use in PCR. PCR of each sample was performed in triplicate for 26 cycles (long mitochondrial and nuclear DNA) and 20 cycles (short mitochondrial DNA), and the PCR products were quantified using the PicoGreen assay. (A and D) long mitochondrial DNA fragments from H99 and BY4741, (B and E) long nuclear DNA fragments from H99 and BY4741, (C and F) short mitochondrial DNA fragments from H99 and BY4741. The solid lines connect all of data points. The dotted lines are the linear regression lines. Results were from three independent experiments with triplicate samples in each experiment (3 samples/point/experiment x 3). Mean ± S.E.M.

    Journal: PLoS ONE

    Article Title: Measuring radiation-induced DNA damage in Cryptococcus neoformans and Saccharomyces cerevisiae using long range quantitative PCR

    doi: 10.1371/journal.pone.0207071

    Figure Lengend Snippet: Quantitative amplification of target DNA fragments in LR-QPCR with respect to the amount of template DNA. DNA isolated from C . neoformans H99 and S . cerevisiae BY4741 were serially diluted with nuclease-free water for use in PCR. PCR of each sample was performed in triplicate for 26 cycles (long mitochondrial and nuclear DNA) and 20 cycles (short mitochondrial DNA), and the PCR products were quantified using the PicoGreen assay. (A and D) long mitochondrial DNA fragments from H99 and BY4741, (B and E) long nuclear DNA fragments from H99 and BY4741, (C and F) short mitochondrial DNA fragments from H99 and BY4741. The solid lines connect all of data points. The dotted lines are the linear regression lines. Results were from three independent experiments with triplicate samples in each experiment (3 samples/point/experiment x 3). Mean ± S.E.M.

    Article Snippet: DNA sample concentrations were determined by fluorescence measurements after DNA staining with PicoGreen (Quant-iT PicoGreen dsDNA reagent and kit; Thermo Fisher Scientific, New York, NY).

    Techniques: Amplification, Real-time Polymerase Chain Reaction, Isolation, Polymerase Chain Reaction, Picogreen Assay

    Murine and human neutrophils release more dsDNA when exposed to L. (V) p . strains that are resistant to either MA or MIL. Bone marrow derived murine neutrophils (A) and blood-derived human neutrophils from healthy donors (B) were exposed to L. (V.) p . of the indicated susceptibility at a 5:1 MOI. As controls, incubations with PMA, PMA + DNase or without stimulus were also performed. Four hours later, NET formation was quantified by measuring the levels of double stranded DNA (dsDNA) released in the supernatant using the PicoGreen fluorescent dye assay. * p

    Journal: Frontiers in Immunology

    Article Title: Resistance of Leishmania (Viannia) Panamensis to Meglumine Antimoniate or Miltefosine Modulates Neutrophil Effector Functions

    doi: 10.3389/fimmu.2018.03040

    Figure Lengend Snippet: Murine and human neutrophils release more dsDNA when exposed to L. (V) p . strains that are resistant to either MA or MIL. Bone marrow derived murine neutrophils (A) and blood-derived human neutrophils from healthy donors (B) were exposed to L. (V.) p . of the indicated susceptibility at a 5:1 MOI. As controls, incubations with PMA, PMA + DNase or without stimulus were also performed. Four hours later, NET formation was quantified by measuring the levels of double stranded DNA (dsDNA) released in the supernatant using the PicoGreen fluorescent dye assay. * p

    Article Snippet: NET formation was also assessed through the measurement of dsDNA in the supernatant using the Quant-iT PicoGreen kit (Thermo Fisher) by adapting a technique previously described ( ).

    Techniques: Derivative Assay

    DNA quantity per scaffold at 3 and 7 days, assessed by PicoGreen assay. This confirms the ability of all scaffold architectures to support primary kidney cell life. Analysis using a one-way ANOVA showed significant differences F(7,23) = 4.79, p = 0.002, post hoc Tukey analysis shows that was in regards to cryogenic scaffolds. Data presented as mean ± 95% confidence intervals, circles denote individual data points

    Journal: Tissue Engineering and Regenerative Medicine

    Article Title: A Non-woven Path: Electrospun Poly(lactic acid) Scaffolds for Kidney Tissue Engineering

    doi: 10.1007/s13770-017-0107-5

    Figure Lengend Snippet: DNA quantity per scaffold at 3 and 7 days, assessed by PicoGreen assay. This confirms the ability of all scaffold architectures to support primary kidney cell life. Analysis using a one-way ANOVA showed significant differences F(7,23) = 4.79, p = 0.002, post hoc Tukey analysis shows that was in regards to cryogenic scaffolds. Data presented as mean ± 95% confidence intervals, circles denote individual data points

    Article Snippet: Total DNA content of the samples was calculated using a Quant-iT™ PicoGreen® assay kit (ThermoFisher, UK) as per the manufacturers’ instructions.

    Techniques: Picogreen Assay

    POLA1 deficiency is associated with reduced cytosolic RNA:DNA levels ( a ) Immunoprecipitation of dsDNA and RNA:DNA from cytosolic fractions of HEK293T cells using the corresponding antibodies and Protein-G agarose beads. Beads were stained with Picogreen ® and visualized by fluorescence microscopy. Nucleic acid recovery was quantified by image analysis (ImageJ). Mean relative fluorescence levels are indicated in parantheses. ( b ) As in (a), dsDNA or RNA:DNA were quantified in control or XLPDR-derived fibroblasts. ( c ) RNA:DNA was quantified in a similar manner in three models of POLA1 deficiency: HEK293T and HeLa after POLA1 silencing (compared to control siRNA), and XLPDR-derived dermal fibroblasts (compared to control cells). Representative images are shown and relative fluorescence levels are indicated. ( d ) Cytosolic RNA:DNA content was quantified in XLPDR-derived fibroblasts stably transduced with a lentivirus encoding HA-tagged wild-type POLA1 (HA-POLA1wt) or EV. Representative images and relative fluorescence levels are indicated. ( e ) Immunoblot analysis of total and p-TBK1, and total and HA-tagged POLA1 expression in XLPDR fibroblast stably expressing HA-POLA1wt or EV (as in d). ( f ) qRT-PCR analysis of IFIT1 mRNA expression in the same cells depicted in (d); when indicated, the cells were treated with poly(dA:dT) for 16h. Scale bars are 20 μm. *p≤0.05, **p≤0.01, NS – not significant (unpaired Student’s t -test). In ( f ) all comparisons are made against the corresponding WT conditions. Data are representative of 1 ( d,e ), 2 ( a,b,f ), or 4 ( c ) independent experiments (mean ( a,c,d ) and mean and s.e.m.( b,f )).

    Journal: Nature immunology

    Article Title: DNA polymerase-α regulates type I interferon activation through cytosolic RNA:DNA synthesis

    doi: 10.1038/ni.3409

    Figure Lengend Snippet: POLA1 deficiency is associated with reduced cytosolic RNA:DNA levels ( a ) Immunoprecipitation of dsDNA and RNA:DNA from cytosolic fractions of HEK293T cells using the corresponding antibodies and Protein-G agarose beads. Beads were stained with Picogreen ® and visualized by fluorescence microscopy. Nucleic acid recovery was quantified by image analysis (ImageJ). Mean relative fluorescence levels are indicated in parantheses. ( b ) As in (a), dsDNA or RNA:DNA were quantified in control or XLPDR-derived fibroblasts. ( c ) RNA:DNA was quantified in a similar manner in three models of POLA1 deficiency: HEK293T and HeLa after POLA1 silencing (compared to control siRNA), and XLPDR-derived dermal fibroblasts (compared to control cells). Representative images are shown and relative fluorescence levels are indicated. ( d ) Cytosolic RNA:DNA content was quantified in XLPDR-derived fibroblasts stably transduced with a lentivirus encoding HA-tagged wild-type POLA1 (HA-POLA1wt) or EV. Representative images and relative fluorescence levels are indicated. ( e ) Immunoblot analysis of total and p-TBK1, and total and HA-tagged POLA1 expression in XLPDR fibroblast stably expressing HA-POLA1wt or EV (as in d). ( f ) qRT-PCR analysis of IFIT1 mRNA expression in the same cells depicted in (d); when indicated, the cells were treated with poly(dA:dT) for 16h. Scale bars are 20 μm. *p≤0.05, **p≤0.01, NS – not significant (unpaired Student’s t -test). In ( f ) all comparisons are made against the corresponding WT conditions. Data are representative of 1 ( d,e ), 2 ( a,b,f ), or 4 ( c ) independent experiments (mean ( a,c,d ) and mean and s.e.m.( b,f )).

    Article Snippet: The samples were quantified using Quant-iT™ PicoGreen® dsDNA Assay Kit (ThermoFisher Scientific, Grand Island, NY) according to manufacturer’s protocol.

    Techniques: Immunoprecipitation, Staining, Fluorescence, Microscopy, Derivative Assay, Stable Transfection, Transduction, Hemagglutination Assay, Expressing, Quantitative RT-PCR